PLGA from PolySciTech used in development of nanoparticle delivery system for CRISPR–Cas9 plasmid

Although gene-editing has been promoted as a potential usage of CRISPR technology there remains a lack of method to deliver the components for this to the intraceullular area. Recently, researchers at Virginia Tech used PLGA (AP063) and PLGA-NH2 (AI063) from PolySciTech (www.polyscitech.com) to create plasmid-encapsulated nanoparticles and measured the release from these as part of a DNA delivery system. This research holds promise to unlock the potential of CRISPR by allowing the DNA systems to be uptaken into target cells. Read more: Jo, Ami, Veronica M. Ringel-Scaia, Dylan K. McDaniel, Cassidy A. Thomas, Rui Zhang, Judy S. Riffle, Irving C. Allen, and Richey M. Davis. "Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR–Cas9 plasmid." Journal of Nanobiotechnology 18, no. 1 (2020): 16. https://link.springer.com/article/10.1186/s12951-019-0564-1

“Background: The clustered regularly interspaced short palindromic repeats (CRISPR) and Cas9 protein system is a revolutionary tool for gene therapy. Despite promising reports of the utility of CRISPR–Cas9 for in vivo gene editing, a principal problem in implementing this new process is delivery of high molecular weight DNA into cells. Results: Using poly(lactic-co-glycolic acid) (PLGA), a nanoparticle carrier was designed to deliver a model CRISPR–Cas9 plasmid into primary bone marrow derived macrophages. The engineered PLGA-based carriers were approximately 160 nm and fluorescently labeled by encapsulation of the fluorophore 6,13-bis(triisopropylsilylethynyl) pentacene (TIPS pentacene). An amine-end capped PLGA encapsulated 1.6 wt% DNA, with an encapsulation efficiency of 80%. Release studies revealed that most of the DNA was released within the first 24 h and corresponded to ~ 2–3 plasmid copies released per nanoparticle. In vitro experiments conducted with murine bone marrow derived macrophages demonstrated that after 24 h of treatment with the PLGA-encapsulated CRISPR plasmids, the majority of cells were positive for TIPS pentacene and the protein Cas9 was detectable within the cells. Conclusions: In this work, plasmids for the CRISPR–Cas9 system were encapsulated in nanoparticles comprised of PLGA and were shown to induce expression of bacterial Cas9 in murine bone marrow derived macrophages in vitro. These results suggest that this nanoparticle-based plasmid delivery method can be effective for future in vivo applications of the CRISPR–Cas9 system. Keywords: Nanoprecipitation Transfection CRISPR–Cas9 PLGA nanoparticles”


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